ArticleEukaryotic cell2015
Genetic mapping reveals that sinefungin resistance in Toxoplasma gondii is controlled by a putative amino acid transporter locus that can be used as a negative selectable marker.
Article in Eukaryotic cell, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- A transcriptional network required for bradyzoite development in Toxoplasma gondii is dispensable for recrudescent disease.Nature communications · 2023Article
- Mode of action studies confirm on-target engagement of lysyl-tRNA synthetase inhibitor and lead to new selection marker forFrontiers in cellular and infection microbiology · 2023Article
- ThemBio · 2021Article
- Article
- Amino Acid Metabolism in Apicomplexan Parasites.Metabolites · 2021Review
- Generation of Toxoplasma gondii and Hammondia hammondi Oocysts and Purification of Their Sporozoites for Downstream Manipulation.Methods in molecular biology (Clifton, N.J.) · 2020Article
- Genomewide Analysis of Mode of Action of themSystems · 2019Article
- The tyrosine transporter of Toxoplasma gondii is a member of the newly defined apicomplexan amino acid transporter (ApiAT) family.PLoS pathogens · 2019Article
- ThemSphere · 2018Article
- QTL Mapping and CRISPR/Cas9 Editing to Identify a Drug Resistance Gene in Toxoplasma gondii.Journal of visualized experiments : JoVE · 2017Article
- Genome Editing by CRISPR/Cas9: A Game Change in the Genetic Manipulation of Protists.The Journal of eukaryotic microbiology · 2016Review
- Identification of small molecule inhibitors that block theACS infectious diseases · 2016Article
- Toxoplasma gondii Infections Alter GABAergic Synapses and Signaling in the Central Nervous System.mBio · 2015Article
- Rhoptry Proteins ROP5 and ROP18 Are Major Murine Virulence Factors in Genetically Divergent South American Strains of Toxoplasma gondii.PLoS genetics · 2015Article
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3 authors.
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Abstract
Quantitative trait locus (QTL) mapping studies have been integral in identifying and understanding virulence mechanisms in the parasite Toxoplasma gondii. In this study, we interrogated a different phenotype by mapping sinefungin (SNF) resistance in the genetic cross between type 2 ME49-FUDR(r) and type 10 VAND-SNF(r). The genetic map of this cross was generated by whole-genome sequencing of the progeny and subsequent identification of single nucleotide polymorphisms (SNPs) inherited from the parents. Based on this high-density genetic map, we were able to pinpoint the sinefungin resistance phenotype to one significant locus on chromosome IX. Within this locus, a single nonsynonymous SNP (nsSNP) resulting in an early stop codon in the TGVAND_290860 gene was identified, occurring only in the sinefungin-resistant progeny. Using CRISPR/CAS9, we were able to confirm that targeted disruption of TGVAND_290860 renders parasites sinefungin resistant. Because disruption of the SNR1 gene confers resistance, we also show that it can be used as a negative selectable marker to insert either a positive drug selection cassette or a heterologous reporter. These data demonstrate the power of combining classical genetic mapping, whole-genome sequencing, and CRISPR-mediated gene disruption for combined forward and reverse genetic strategies in T. gondii.
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